Use of revitalizing cosmetic composition and non-therapeutic methods

ABSTRACT

The present invention relates to the use of revitalizing cosmetic compositions in association with a light emitting diode (LED) as skin cell stimulators and a related non-therapeutic method for the skin treatment of aging and for repair and recovery of cutaneous tissue.

FIELD OF THE INVENTION

This invention relates to the use of revitalizing cosmetic compositionsin association with a light emitting diode (LED) as skin cellstimulators and a related non-therapeutic method for the skin treatmentof aging and for repair and recovery of cutaneous tissue.

BACKGROUND OF THE INVENTION

The exposure of the skin to a variety of agents causes its extrinsicaging. These agents can be any of the following causes: pollution,genetic dispositions, solar radiation and UV exposure, among others. Inaddition to external factors, internal agents are responsible forintrinsic aging. Skin aging can be defined as a fall and/or decay of therate of the metabolism efficiency due to insufficient production ofsubstances and cells to promote tissue regeneration. Moreover, due tothe loss of elasticity and firmness and consequent fragility, the skinaging can be characterized as being intrinsic when there is theidentification of a superficial wrinkling, as well as thinning andsagging of the skin, or being extrinsic, when the presence of deepwrinkles and pockets, dryness, and pigmentation irregularities is noted(Emerit, I. “Free Radicals and Aging of the Skin”. Birkhäuser Basel1992; 62:328-341).

In order to solve the problem of skin aging, various non-therapeutictechniques and forms of non-therapeutic treatment are suggested in theliterature and by another important segment, the cosmetic industry,which promotes treatments using active ingredients incorporated intocreams, cosmetic compositions, gels, elixirs among others.

In addition, it is considered that the associated use of differenttreatment methods, such as cosmetics and chemicals, has a trend toprovide a significant improvement in the skin, contributing to itstreatment and rejuvenation.

There are several prior art documents relating to methods andcompositions used to improve and/or treat skin aging. Among them,reference may be made to ZA1998/00404, which relates to a cosmetic skintreatment composition comprising from 0.1% to 75% of at least onecellular therapeutic compound, substance or composition, that isselected from hyaluronic acid or a pharmaceutically acceptable saltthereof, a keratin binding complex, a collagen amino acid, a compositionincluding sericin and glycoprotein, among others, in addition to acosmetically acceptable carrier or diluent. Further it discloses asubstance or composition for use in a method of treating the human skin,wherein in said method there is the application to the skin of aneffective amount of the substance or composition with subsequentincidence of laser irradiation to the subject's skin who needstreatment.

The document US20130115180 relates to compositions for administeringelectromagnetic radiation (EMR) for therapeutic or cosmetic purposes andmethods, wherein said compositions comprise a carrier lotion of activecosmetic ingredients which is suitable for application to human skin; afirst and second compositions suitable for application to the humanskin, wherein the first preferably blocks the EMR below a certainpredetermined wavelength range and the second preferably blocks the EMRabove another predetermined wavelength range; and the source of lightused may be artificial or ultraviolet light.

Thus, it is still desirable to find topically applicable compositionsfor skin treatment which, when exposed to a particular type ofradiation, especially radiation from light emitting diode (LED), areable to show significant improvement results in relation to the issue ofskin aging and in the recovery of cutaneous tissue, more specificallythat they present cellular stimulation, and consequently, reduction ofthe aging signs and also, the repair of cutaneous tissue.

SUMMARY OF THE INVENTION

As previously mentioned, this invention relates to the use ofrevitalizing cosmetic compositions which by photobiomodulation (the termrepresenting cell stimulation from a light-emitting diode) on certainLED wavelengths, amplify the synthesis of certain essential proteins andexpressed in fibroblast culture, such as collagen I, collagen III,fibronectin, laminin, α-SMA.

A preferred embodiment of the invention refers to the use of therevitalizing cosmetic compositions, particularly elixirs comprisinggrowth factors, platinum, diamond and gold particles-conjugatespeptides, pearl extract and caviar hydrolysate in combination with LEDfor optimization of the synthesis of cellular proteins selected from thegroup consisting of collagen I, collagen III, fibronectin, laminin andα-SMA.

In another preferred embodiment of the invention, there is disclosed anon-therapeutic method for the skin aging treatment, said methodcomprising combining revitalizing cosmetic compositions, in particularelixirs comprising growth factors, platinum, diamond and goldparticles-conjugated peptides, pearl extract and caviar hydrolysate,with LEDs at certain wavelengths.

In yet another embodiment of the invention, there is disclosed anon-therapeutic method for repairing and recovering cutaneous tissue,said method also comprising the association of revitalizing cosmeticcompositions, in particular elixirs comprising growth factors, platinum,diamond and gold particles-conjugated peptides, pearl extract and caviarhydrolysate, with LEDs at certain wavelengths.

Unexpectedly, the inventors of this invention have noted a synergisticpotentiating action by associating the revitalizing cosmeticcompositions with the wavelength lighting determined in the visible/LEDlight spectrum, continuously emitted over a given time. The subjectmatter of the invention is therefore a novel non-therapeutic method ofskin aging treatment. It is a method that combines “in vitro” light(photobiomodulation) and revitalizing cosmetic compositions(bioinduction) for repair and recovery of cutaneous tissue, in which aset of induced cellular proteins is measured against the appliedlighting—expressing a cellular metabolic state that determines newregeneration capacities.

These features of the invention will be described in more detail in thedetailed description of the invention.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows qualitatively the increase in laminin expression due to thesynergistic association of the use of 1% of the gold elixir and exposureto the various LED wavelengths.

FIG. 2 shows qualitatively the increase in collagen I and III expressiondue to the synergistic association of the use of 1% of the diamondelixir and exposure to the various LED wavelengths.

FIG. 3 shows qualitatively the increase of α-SMA expression as a resultof the synergistic association of the use of 1% of the diamond elixirand exposure to the various LED wavelengths.

FIG. 4 shows qualitatively the increase in fibronectin expression due tothe synergistic association of the use of 1% of the gold elixir ordiamond elixir and exposure to the various LED wavelengths.

FIGS. 5 to 10 show graphs where the structural proteins andextracellular matrix elements are quantified.

FIGS. 11 to 12 show the migration tests performed for fibroblasts.

DETAILED DESCRIPTION OF THE INVENTION

It is presented in this description the use of revitalizing cosmeticcompositions in association to LED to optimize the extracellular matrixprotein synthesis, and further the non-therapeutic method for skin agingtreatment and repairing and recovering cutaneous tissue, said methodscomprising the association of revitalizing cosmetic compositions, inparticular elixirs comprising growth factors, platinum, diamond and goldparticles-conjugated peptides, pearl extract and caviar hydrolysate,with LEDs at certain wavelengths.

By associating the revitalizing cosmetic compositions with the lightemitting diode (LED), and evaluating the synthesis of structuralproteins, the inventors surprisingly observed a potentiating synergisticeffect of the association of the cosmetic/light compositions on theculture of fibroblasts compared to cultures of cells maintained in theabsence of illumination.

The revitalizing cosmetic compositions of this invention are formulatedas follows:

Pearl and Caviar Elixir: formulated to hydrate, inhibit tyrosinase andrevitalize the skin. The high content of proteins and vitamins frompearl and caviar extracts activates new skin cells gradually reducingthe formation of wrinkles, in addition to hydrating and helping toprevent loss of skin elasticity, promoting a more uniform texture.

Colloidal Gold Elixir: elixir specially formulated to remineralize theskin layers, with antioxidant and stimulus action to the synthesis ofstructural proteins of the skin, such as collagen (collagen I and III)and elastin. It stimulates healing, fibroblasts, filling and conductionof bioelectricity. The peptide conjugated with modified goldnanomaterial hydrates, revitalizes and unifies the texture of thecutaneous tissue, conferring to it, softness and luminosity.

Platinum Elixir: elixir specially formulated to restore the skin barrierby maintaining epithelial thickness through hyaluronic acid, GAGstructural proteins and Langerhans cells, reducing the dermal-epidermalcell matrix degradation and acting on healing. The enzyme peptideconjugated with platinum nanomaterial enhances the production ofstructural proteins, hydrates and protects the skin againstenvironmental stress, conferring vitality and protection.

Diamond Elixir—elixir specially formulated to reshape and rebuild skinstructures through collagen VII, laminin-5, fibronectin, reducingwrinkles and expression lines, improving healing and filling. The highconcentration of available peptide with diamond nanomaterial hydratesand helps preventing loss of skin elasticity, giving young and healthyappearance with renewed texture.

The structural proteins expressed in the extracellular matrix areselected from the group consisting of: collagen I, collagen III,fibronectin, laminin and α-SMA

The non-therapeutic method for the skin aging treatment of thisinvention therefore comprises the following steps:

-   -   a) applying an acceptable amount of at least one revitalizing        cosmetic composition onto the cutaneous tissue;    -   b) subjecting the revitalizing cosmetic composition applied        in (a) light having wavelengths between 410 nm and 780 nm;    -   c) maintaining the exposure of the revitalizing cosmetic        composition to light as described in (b) for a time period        ranging from at least 15 seconds to 2 minutes.

The non-therapeutic method for repairing and recovering cutaneous tissuecomprises the following steps:

-   -   a) applying an acceptable amount of at least one revitalizing        cosmetic composition onto the injured or discontinued cutaneous        tissue; 0.35 to 70 ml    -   b) subjecting the revitalizing cosmetic composition applied        in (a) light having wavelengths between 410 nm and 780 nm;    -   c) maintaining the exposure of the revitalizing cosmetic        composition as described in (b) for a time period ranging from 5        hours to 40 hours, more preferably, between 20 hours to 40        hours.

In a preferred embodiment of the invention the amount of therevitalizing cosmetic composition ranges from 0.35 ml to 15 ml, morepreferably the amount of the revitalizing cosmetic composition to beapplied to the skin tissue is from 0.35 ml to 1.50 ml.

The inventors when measuring cutaneous markers/cellular proteins from acontrol sample where only the revitalizing cosmetic composition wasapplied and compared with samples that received the revitalizingcosmetic composition and were also subjected to LED light withwavelengths ranging from 410 nm and 780 nm, surprisingly obtained theresults as shown in Tables 1 to 5 as follows:

TABLE 1 Collagen 1 ELIXIR 1% Sem LUZ 410 nm 440 nm 470 nm 520 nm 590 nm630 nm 660 nm 690 nm 780 nm 470-850 nm CTRL * Pearl & Cavlar * * * ** *** * * ** * * Gold * ** ** ** * ** ** * ** * Platin * * * ** * ***** * * * * Diamond ** ** ** ** ** *** *** *** * ** *

TABLE 2 Collagen III ELIXIR 1% Sem LUZ 410 nm 440 nm 470 nm 520 nm 590nm 630 nm 660 nm 690 nm 780 nm 470-850 nm CTRL Pearl &Cavlar * * * * * * * * * * * Gold * * * * * * * * * * *Platin * * * * * * * * * * * Diamond * ** ** ** ** *** *** ** ** ** ***

TABLE 3 fibronectin ELIXIR 1% Sem LUZ 410 nm 440 nm 470 nm 520 nm 590 nm630 nm 660 nm 690 nm 780 nm 470-850 nm CTRL * Pearl & Cavlar * * * * * *** * ** * * Gold ** ** ** *** *** *** *** ** * * ** Platin * * * * * * *** ** ** * Diamond ** ** *** *** *** *** *** *** ** ** *

TABLE 4 αSMA (myofibroblasts) ELIXIR 0.15% Sem LUZ 410 nm 440 nm 470 nm520 nm 590 nm 630 nm 660 nm 690 nm 780 nm 470-850 nm CTRL * Pearl &Cavlar * * ** ** ** * ** ** * / / Gold * * ** * * ** ** ** ** / / Platin** ** ** ** ** *** ** ** ** *** / Diamond * ** ** ** ** *** ** * * / /

TABLE 5 Laminin ELIXIR 1% Sem LUZ 410 nm 440 nm 470 nm 520 nm 590 nm 630nm 660 nm 690 nm 780 nm 470-850 nm CTRL * Pearl &Cavlar * * * * * * * * * * / Gold * * ** *** *** ** * ** ** ** **Platin * * * * * * * * * * / Diamond * * * * * * * * * * /* A red dot corresponding to the control or base level. Two red dotsindicate that the skin marker production was moderately larger than thatof the control. Three red dots indicate that the production of the skinmarker was significantly greater than that of the control.

According to this invention, the terms “revitalizing cosmeticcompositions” and “elixirs” are used interchangeably and refer tocompositions for treating the aging signs.

Furthermore, the terms “skin markers” and “certain essential proteins”are used interchangeably and refer to proteins synthesized byfibroblasts, and make up the extracellular matrix.

The revitalizing cosmetic compositions or elixirs are formulated withexcipients, humectants, preservatives, opacifier, thickener, skinprotectant, conditioner, colorant, emollient, chelating, emulsifier, pHadjuster, fragrance, growth factor, and further, vehicle options.

Cosmetically acceptable excipients include, without limitation, pHadjusting agents, conditioners, preservatives, colorants, emollients,emulsifiers, fragrances, thickeners, sequestering and carriers.

Examples of pH adjusting agents include, without limitation, aceticacid, citric acid, hydrochloric acid, lactic acid, sodium bicarbonate,ammonium carbonate, potassium hydroxide, sodium hydroxide,triethanolamine.

Examples of conditioners include, without limitation, copperacetylmethionate, magnesium acetylmethionate, manganeseacetylmethionate, zinc acetylmethionate, methylsilanol hydroxyprolineaspartate, carnosine and its derivatives, plant extracts such as oliveleaf extract (Olea europaea), glutamylamidoethyl imidazole, vegetableoils such as cottonseed oil (Gossypium herbaceum), sunflower seed oil(Helianthus annuus), passion fruit seed oil (Passiflora edulis), grapeseed oil (Vitis vinifera).

Examples of preservatives include, without limitation, benzoic acid,benzyl alcohol, sodium benzoate, cetylpyridinium chloride, benzalkoniumchloride, benzethonium chloride, phenoxyethanol, imidazolinidyl urea,parabens and mixtures thereof.

Examples of colorants include, without limitation, CI10315 (yellow),CI12085 (red), CI15510 (orange), CI15800 (red), CI15880 (red), CI15985(yellow), CI19140 (yellow), CI20170 (brown), CI42053 (green), CI42080(blue), CI42090 (blue), CI42510 (violet), CI45100 (red), CI45370(orange), CI59040 (green), CI60725 (violet), CI60730 (violet), CI75170(white), caramel and mixtures thereof.

Examples of emollients include, without limitation, stearic acid, lacticacid, animal fats such as lanolin, vegetable oils such as cottonseed oil(Gossypium herbaceum), sunflower seed oil (Helianthus annuus), passionfruit seed oil (Passiflora edulis), grape seed oil (Vitis vinifera),propylheptyl caprylate, caprylyl glycol, coco caprylate/caprate,cyclomethicone, dimethicone, ethoxydiglycol, glycerin, lactose, cetylpalmitate, sorbitol, caprylic/capric acid triglyceride, urea.

Examples of emulsifiers include, without limitation, cetearyl alcohol,cetyl alcohol, stearyl alcohol and mixtures thereof, glyceryl stearate,ethoxylated fatty alcohols, sorbitan esters such as sorbitan oleate andsorbitan stearate, lecithin and polysorbates.

Examples of thickeners include, without limitation, waxes, such asbeeswax, carnauba wax and lanolin wax and lanolin, polysaccharides,among which starch, gums such as arabic gum, guar gum, xanthan gum,tragacanth, agar, carrageenans and alginates, cellulose and itsderivatives, such as microcrystalline cellulose, cellulose acetate,carboxymethylcellulose and hydroxyethylcellulose, glyceryl stearate,polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, carbopol,polyacrylic acid, silanes and derivatives, alkyl polyacrylates, alkylpolymethacrylates and mixtures thereof.

Examples of fragrances include, without limitation, natural, syntheticfragrances and mixtures thereof.

Examples of sequestrants include, without limitation, EDTA, disodiumEDTA, tetrasodium EDTA, monobasic sodium phosphate and dibasic sodiumphosphate and mixtures thereof.

Examples of carriers include, without limitation, water, esters ingeneral, mineral oil and vegetable oils and mixtures thereof, when intoan emulsified system.

As previously stated, control and exposure of cell culture withrevitalizing cosmetic composition associated with LED stimulation atvarious wavelengths were compared and optimal wavelength ranges wereobtained for the synergistic potentiation of elixir application withexposure to LED for each marker. A synergistic potentiating action wassurprisingly detected in this associated use of revitalizing cosmeticcompositions with LEDs, which provided an increase in the synthesis ofcutaneous markers such as collagen I, collagen III, fibronectin, lamininand α-SMA.

EXAMPLES Evaluations and Results

The methodology for measuring photobiostimulation was to compare thecontrol and exposure of the cell culture with the product afterstimulation of the LED at various wavelengths.

The analyzed wavelengths are in the range from 410 nm to 780 nm.

Non-Therapeutic Method for the Cutaneous Aging Treatment:

Complementing the results of cell stimulation on fibroblasts fromcertain LED wavelengths, evaluation of cell migration on fibroblasts wasdetermined using the associations between the parameters (concentrationof cosmetic composition used, wavelength, time of exposure to LED, forexample).

The fibroblast cell culture was submitted to LED and contact with theelixirs according to the best parameters of the qualitative evaluation(data from tables 1 to 5).

In relation to the method for repair and recovery of cutaneous tissue,the evaluation of the cellular migration effect is demonstrated by thespeed with which the cells repair the artificial discontinuity of thecreated layer (wound maker) to demonstrate the effect that therevitalizing cosmetic compositions, specifically the Gold ColloidalElixir and Diamond Elixir, present in potentiating the healing andrecolonization of injury areas (FIGS. 11 to 12).

FIG. 1 shows qualitative microscopic images of increased lamininexpression as a result of the synergistic potentiation of 1% applicationof the Gold Colloidal Elixir and LED exposure. The comparison wasperformed with control or only with the use of said elixir. Wherein:

-   -   Green: Specific marker signaling.    -   Red: Linking of phalloidin that highlights the cell actin        filaments.    -   Blue: Identification of the cell nucleus.

The qualitative evaluation of Table 5 shows, in a relative way, how theLED wavelengths at 470 and 520 nm in contact with the Gold ColloidalElixir potentiate laminin production by the fibroblasts. Data that isreinforced by the quantitative evaluation of FIG. 9, specifically at 470nm, where exposure to the LED is maintained for 30 seconds.

FIG. 2 shows qualitative microscopic captures of increased type I andIII Collagen expression as a result of the synergistic potentiation of1% application of the Diamond Elixir and LED exposure. The comparisonwas performed with control or only with the use of said elixir. Wherein:

-   -   Green: Specific marker signaling.    -   Red: Linking of phalloidin that highlights the cell actin        filaments.    -   Blue: Identification of the cell nucleus.

The qualitative evaluation of Tables 1 and 2 shows, in a relative way,how the LED wavelengths at 590, 630 and 660 nm in contact with theDiamond Elixir potentiate type I and III Collagen production by thefibroblasts. Data that are reinforced by the quantitative evaluation ofFIGS. 5 and 6, specifically at 470 nm where the LED exposure ismaintained for 30 seconds for type I collagen production and 590 nmwhere the exposure to the LED is maintained for 15 seconds forproduction of type III collagen.

FIG. 3 shows qualitative microscopic images of increased α-SMAexpression as a result of the synergistic potentiation of 1% applicationof the Diamond Elixir and LED exposure. The comparison was performedwith control or only with the use of said elixir. Wherein:

-   -   Green: Specific marker signaling.

The qualitative evaluation of Table 4 shows, in a relative way, how theLED wavelengths at 590 nm in contact with the Diamond Elixir potentiatethe manifestation of α-SMA marker by the fibroblasts. Data that isreinforced by the quantitative evaluation of FIG. 10, specifically at625 and 660 nm, where exposure to the LED is maintained for 30 seconds.

FIG. 4 shows qualitative microscopic images of increased fibronectinexpression as a result of the synergistic potentiation of 1% applicationof the Gold Elixir or Diamond Elixir and LED exposure. The comparisonwas performed with control or only with the use of said elixir. Wherein:

-   -   Green: Specific marker signaling.

The qualitative evaluation of Table 3 shows, in a relative way, how theLED wavelengths at 470, 520, 590 and 630 nm in contact with the GoldColloidal Elixir potentiate fibronectin production by the fibroblasts.Data that is reinforced by the quantitative evaluation of FIG. 7,specifically at 590 nm, where exposure to the LED is maintained for 30seconds.

The qualitative evaluation of Table 3 shows, in a relative way, how theLED wavelengths at 440, 470, 520, 590, 630 and 660 nm in contact withthe Diamond Elixir potentiate fibronectin production by the fibroblasts.Data that is reinforced by the quantitative evaluation of FIG. 8,specifically at 630 and 590 nm, where exposure to the LED is maintainedfor 30 seconds.

Non-Therapeutic Method for Cutaneous Tissue Repair:

The fibroblast cell culture was submitted to LED and contact with theelixirs according to the best parameters of the qualitative evaluation(data from tables 1 to 5).

The evaluation of the cellular migration effect is demonstrated by thespeed with which the cells repair the artificial discontinuity of thecreated layer (wound maker) to demonstrate the effect that therevitalizing cosmetic compositions, specifically the Gold ColloidalElixir and Diamond Elixir, present in potentiating the healing andrecolonization of injury areas.

Thus, FIGS. 11 and 12 depict the migration tests plots performed forfibroblasts.

FIG. 11 shows the results of fibroblast cell migration on differentconcentrations of the Diamond Elixir, identifying that theconcentrations evaluated between 0.05% and 0.1% presented stimulus closeto the control stimulated only by exposure to the LED.

FIG. 12 shows the results of fibroblast cellular migration on differentconcentrations of the Colloidal Gold Elixir, indicating that theconcentrations evaluated between 1% and 0.5% presented a stimulus higherthan the control stimulated only by exposure to the LED.

On a lower proportion and less potentiating effect when compared to theother results, but not less important, the association between LEDexposure and Pearl & Caviar Elixir, increased the production of type Icollagen in fibroblasts when exposed to 470, 590 and 690 nm. Themanifestation of the α-SMA marker by fibroblasts was evidenced whenexposed to 440, 470, 520, 630 and 660 nm. Fibronectin production infibroblasts is higher when the Pearl & caviar Elixir is exposed at 630and 690 nm.

On a lower proportion and less potentiating effect when compared to theother results, but not less important, the association between LEDexposure and Platinum Elixir, increased the production of type Icollagen in fibroblasts when exposed to 590 nm. The manifestation of theα-SMA marker by fibroblasts was evidenced when exposed to 590 and 780nm. Fibronectin production in fibroblasts is higher when the PlatinumElixir is exposed at 660, 690 and 780 nm.

From the above description and example, it is possible to observe anincrease in the synthesis of cellular proteins, or even in the proteinssynthesized by the fibroblasts, proving an unexpected synergisticpotentiating effect of the use of the revitalizing cosmetic compositionswith LED exposure in relation to the state of the art.

Although certain embodiments have been specifically described, they havebeen presented only by way of example, and there is no intention tolimit the scope of the invention. The claims accompanying thisdisclosure and its equivalents are considered to encompass suchembodiments.

Finally, modifications of this invention apparent to a person skilled inthe art, such as addition or removal of non-fundamental elementsthereof, may be performed without departing from the scope and spirit ofthe invention.

1-2. (canceled)
 3. A method for the skin aging treatment, comprising: a.applying an acceptable amount of at least one revitalizing cosmeticcomposition onto the cutaneous tissue; b. subjecting the revitalizingcosmetic composition applied in (a) to light having wavelengths between410 nm and 780 nm; c. maintaining the exposure of the revitalizingcosmetic composition to light as described in (b) for a time periodranging from at least 15 seconds to 2 minutes.
 4. A method for repairingand recovering cutaneous tissue, comprising: a. applying an acceptableamount of at least one revitalizing cosmetic composition onto injured ordiscontinued cutaneous tissue; b. subjecting the revitalizing cosmeticcomposition applied in (a) to light having wavelengths between 410 nmand 780 nm; c. maintaining the exposure of the revitalizing cosmeticcomposition as described in (b) for a time period ranging from 5 hoursto 40 hours.
 5. The method according to claim 3, wherein the skinmarkers are selected from the group consisting of: collagen I, collagenIII, fibronectin, laminin and α-SMA.
 6. The method according to claim 3,wherein the amount of the revitalizing cosmetic composition to beapplied in cutaneous tissue varies between 0.35 and 15 ml.
 7. The methodaccording to claim 3, wherein the amount of the revitalizing cosmeticcomposition is still more preferably from 0.35 ml to 1.5 ml.
 8. Themethod according to claim 4, wherein the amount of the revitalizingcosmetic composition to be applied in cutaneous tissue varies between0.35 and 15 ml.
 9. The method according to claim 4, wherein the amountof the revitalizing cosmetic composition is from 0.35 ml to 1.5 ml. 10.A method for the skin aging treatment, comprising: a. applying anacceptable amount of at least one revitalizing cosmetic composition ontocutaneous tissue; and b. subjecting the revitalizing cosmeticcomposition applied in (a) to light having wavelengths between 410 nmand 780 nm.